BIOCHEMICAL STUDIES. 243 



No. 10. 1 c.c. 1 per cent venom solution, 10 c.c. olive oil, 0.5 c.c. of 0.8 percent 

 magnesium sulphate solution, 0.5 c.c. water, 3 drops toluol. Increased 

 acidity above No. 9 after incubation = 1 c.c. N/ 10 potassium hydroxid. 



No. 11. 1 c.c. 1 per cent venom solution, 10 c.c. olive oil, 0.5 c.c. of 0.2.sodium 

 taurocholate solution, 0.5 c.c. water, 3 drops toluol. Increased acid- 

 ity above No. 9 after incubation = 0.7 c.c. N/10 potassium hydroxid. 

 Evidently the lipolysis is far less for olive oil than for lecithin. 



No. 12. 1 c.c. water, 10 c.c. 2 per cent lecithin emulsion, 1 c.c. N/10 hydro- 

 chloric acid, three drops toluol. After incubation acidity = 13.5 c.c. 



No. 13. 1 c.c. 1 per cent venom solution, 10 c.c. 2 per cent lecithin emulsion, 

 1 c.c. N/10 hydrochloric acid, 3 drops[toluol. The acidity after incu- 

 bation was the same as No. 12. 



Evidently lecithin is very easily hydrolyzed by mineral acid, and this 

 hydrolysis is not influenced by the venom. It may be that the ease with which 

 lecithin is hydrolyzed by acid accounts for .some of the results obtained in the 

 course of this investigation and by other investigators. It may be one of the 

 factors upon which the greater lipolysis of lecithin depends, for the small quan- 

 tities of acids liberated at the beginning by enzyme action probably tend even 

 without the further help of any enzj'me to hydrolyze the lecithin further. 



No. 14. 1 c.c. water, 10 c.c. 2 per cent lecithin emulsion, 1 c.c. N/10 potassium 

 hydroxid, 3 drops toluol. After incubation acidity = 1.9 c.c. N/10 

 potassium hydroxid. 



No. 15. 1 c.c. 1 per cent venom solution, 10 c.c. 2 per cent lecithin emulsion, 

 1 c.c. N/10 potassium hydroxid, 3 drops toluol. After incubation, 

 acidity after subtracting the control No. 14= 1.6 c.c. N 10 potassium 

 hj'droxid. 



Evidently, while the venom lipolyzes in a solution of this alkalinity, it is 

 more active in less alkaline ones. 



Since these experiments demonstrated that too great an acidity had an 

 action independent of the enzyme, and since too great alkalinity diminished 

 lipolysis, a further series of experiments was undertaken under conditions which 

 should insure continued neutrality in the solution. To attain this end, a solu- 

 tion of 9 gm. disodic phosphate (Na^HPO.,) and 1 gm. of monosodic phosphate 

 (NaH2P04) in 100 c.c. of water was employed, as recommended by L. J. Hen- 

 derson. At the same time the destructive effect of high temperatures was 

 confirmed. The following experiments were set up: 



No. 16. 1 c.c. 1 per cent venom solution, 10 c.c. 2 per cent lecithin emulsion, 

 1 c.c. water, 3 drops toluol. This was titrated at once. Acidity = 

 2.4 c.c. 



No. 17. 1 c.c. 1 per cent venom solution, 10 c.c. 2 per cent lecithin emulsion, 



1 c.c. water, 3 drops toluol. Acidity after incubating and subtracting 

 control No. 16 = 1.2 c.c. It is not clear why in this experiment a 

 lower value was obtained than in No. 3. 



No. 18. 1 c.c. 1 per cent venom solution, 10 c.c. 2 per cent lecithin emulsion, 



2 drops toluol, 1 c.c. phosphate solution. In orderthat the pho.sphate 

 might not interfere with the titration, this solution was exhausted at 

 once with ether, the ether evaporated with an air current, the residue 

 dissolved in alcohol and titrated. Acidity = 2.1 c.c. N 10 potassium 

 hydroxid. 



No. 19. This experiment was in every way identical with No. 18, except that 

 the mixture was incubated before extraction and titration. Acidity 

 = 3.7 c.c. N/10 potassium hydroxid or, subtracting control (No. 18), 

 1.6 c.c. 



